Bacterial chaperones increase the production of soluble human TNF-alpha in Escherichia coli |
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Authors: | Woojin Jeong Nam-Kyu Shin Hang-Cheol Shin |
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Affiliation: | (1) Protein Engineering Laboratory, Hanhyo Institute of Technology, 461-6 Chonmin-dong, Yusong-gu Taejon, 305-390, Republic of Korea |
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Abstract: | Bacterial chaperones were coexpressed to enhance the production of soluble human TNF-alpha and its low-toxicity mutant in Escherichia coli. In the absence of chaperones, about 65% of wild-type TNF and 35% of mutant TNF having a deletion of N-terminal 7 amino acids and substitutions of Leu29'Ser, Ser52;Ile and Tyr56'Phe, were produced as a soluble form. In the presence of overproduced chaperones, most of wild-type and mutant TNF (>95%) were produced as a soluble form, indicating that GroEL and GroES chaperones promote folding and assembly of trimeric TNF-alpha. Bacterial chaperones could be useful in the production of TNF-alpha and its variants as well as other proteins with biological importance. |
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